Establishment of high-throughput-screening model for IRE1 kinase inhibitor

Abstract

Abstract: This paper expressed and purified the cytosolic fragment of human IRE1 （inositol-requiring enzyme1）, an ER transmembrane 'sensor〖KG-*9〗' protein with bacular expression system. Then the HTRF (Homogeneous Time-resolved Fluorescence) assay for IRE1 kinase was set up and optimized. Z′-factor is about 0.58 and the coefficients of variation (CV) is less than 10%, indicating the molecue-based high-throughput screening assay for IRE1 kinase inhibitor was successfully established. This work would lay a solid foundation for the discovery of IRE1 kinase inhibitor and the further research.

Key words: IRE1, kinase assay, bacular expression, HTRF, high-throughput screening

CLC Number:

ZHANG Kun-zhi， SU Ming-bo， ZHOU Yu-bo. Establishment of high-throughput-screening model for IRE1 kinase inhibitor[J]. Journal of East China Normal University(Natural Sc, 20120, 2012(6): 103-113.

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http://xblk.ecnu.edu.cn/EN/Y20120/V2012/I6/103

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